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. 2020 Jun 10;9(11):e016099. doi: 10.1161/JAHA.120.016099

Figure 2. Deletion of Nck 1 but not Nck2 adaptor protein blunts shear stress–induced endothelial permeability.

Figure 2

A, Representative blots showing the efficiency of the knocking down of Nck1/2 in human aortic endothelial cells (HAECs) after Nck1 short hairpin RNA (shRNA), Nck2 shRNA, or scrambled shRNA control transduction. B and C, PECAM‐1 (platelet endothelial cell adhesion molecule‐1; white)/ß‐catenin (red)–stained cells showing paracellular formation (white arrowheads) after oscillatory shear stress (OSS) exposure. D and E, Nck1 but not Nck2 knocking down prevents flow‐induced permeability using biotinylated gelatin/Alexa 647‐streptavidin assay. 4′,6‐Diamidino‐2‐phenylindole (blue; nuclei), phalloidin (green; F‐actin fibers), and streptavidin (red; exposed biotinylated gelatin). Images were analyzed using NIS‐Elements software. Scale bars=50 to 100 μm. Data are from n=4 and analyzed as mean±SEM by 2‐way ANOVA and Bonferroni posttest (*P<0.05, **P<0.01, ***P<0.001). Images in the boxes indicate areas of higher magnifications, whereas areas in the dashed boxes indicate lower magnifications.